EFFECT OF SITE-DIRECTED MUTAGENESIS ON CONSERVED POSITIONS OF DROSOPHILA ALCOHOL-DEHYDROGENASE

被引：49

作者：

COLS, N ^{[1
]}

MARFANY, G ^{[1
]}

ATRIAN, S ^{[1
]}

GONZALEZDUARTE, R ^{[1
]}

机构：

[1] UNIV BARCELONA,FAC BIOL,DEPT GENET,AV DIAGONAL 645,E-08028 BARCELONA,SPAIN

来源：

FEBS LETTERS | 1993年 / 319卷 / 1-2期

关键词：

SITE-DIRECTED MUTAGENESIS; ACTIVE SITE RESIDUE; ALCOHOL DEHYDROGENASE; SHORT-CHAIN DEHYDROGENASE; DROSOPHILA;

D O I：

10.1016/0014-5793(93)80043-T

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Tyr152 and Lys156 may be functionally important residues in Drosophila ADH as they are conserved in the genus and in all short-chain dehydrogenases. In addition, unaltered Gly positions could have a crucial role in the building of the structural framework. We have modified Drosophila ADH and expressed the mutant forms in E. coli. Mutation of Tyr152 to Glu or Gln, Lys156 to Ile, Gly184 to Leu, and the double mutant Gly130 to Cys and Gly133 to Ile, all rendered, with different substrates and at different pHs, an inactive enzyme. Results suggest that Tyr152 and Lys156 are involved in catalysis and that Gly130, Gly133 and Gly184 contribute substantially to the structure of the active form.

引用

页码：90 / 94

页数：5

共 22 条

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